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Diaphot 300 inverted

Manufactured by Nikon
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The Nikon Diaphot 300 inverted microscope is a versatile laboratory instrument designed for a range of microscopy applications. Its inverted configuration allows for the observation of samples from below, making it suitable for cell culture, tissue analysis, and other biological research applications. The Diaphot 300 features high-quality optics and a stable, ergonomic design to provide users with a reliable and efficient tool for their laboratory work.

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Lab products found in correlation

Acetate and tartrate solution, by Merck Group (1 mentions) Microplate reader, by BMG Labtech (1 mentions) P nitrophenol phosphate, by Merck Group (1 mentions) Trap activity, by Merck Group (1 mentions) Poly l lysine solution, by Merck Group (1 mentions) Hq510lp, by Chroma Technology (1 mentions) 505dcxru, by Chroma Technology (1 mentions) Pdmi 2, by Harvard Apparatus (1 mentions) Fluorescein in situ cell death detection kit, by Roche (1 mentions)

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Diaphot 300 (111 citations) Diaphot (102 citations) Diaphot 300 inverted microscope (6 citations)

3 protocols using diaphot 300 inverted

1

Quantification of Osteoclast TRAP Activity

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Osteoclasts on glass coverslips were fixed with 4% paraformaldehyde, and stained for TRAP activity (Sigma) according to the manufacturer's manual. Images were captured using a Nikon Diaphot 300 inverted microscope. TRAP activity was quantified by a colorimetric method. Cells grown in 48-well plates were washed with Hanks Buffer and incubated at 37°C with a pre-warmed, 200 μL mixture containing 0.1% SDS, 2 mg p-nitrophenol phosphate, acetate and tartrate solution (Sigma) for 30 min. The reaction was stopped by adding 40 μL of 0.5 M NaOH and absorbance was read at 405 nm in a microplate reader (BMG Labtech, Cary, NC).
Sharma R., Callaway D., Vanegas D., Bendele M., Lopez-Cruzan M., Horn D., Guda T., Fajardo R., Abboud-Werner S, & Herman B. (2014). Caspase-2 Maintains Bone Homeostasis by Inducing Apoptosis of Oxidatively-Damaged Osteoclasts. PLoS ONE, 9(4), e93696.
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2

Fluorescence Imaging of Labeled Spermatozoa

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Fluorescence imaging was carried out as previously reported (Nishigaki et al., 2004 (link)). Briefly, Fluo-4-labeled spermatozoa were adhered to glass coverslips coated with 50 Pg/ml poly-L-lysine solution (Sigma) and mounted into a micro incubator, PDMI-2 (Harvard Apparatus, Holliston, MA, USA) maintained at 14 °C. Fluorescence images were acquired using a Nikon DIAPHOT 300 inverted microscope with a Nikon Plan Apo 60X objective lens (1.4 NA) and a custom-built stroboscopic illumination system (Nishigaki et al., 2006 (link)) with a Chroma filter set (Ex, HQ470/40x; DC, 505DCXRU; Em, HQ510LP (Chroma Technology)). Images were acquired with a Quantix 57 camera (Photometrics Inc.) under the continuous (stream) acquisition mode.
Beltrán C., Rodríguez-Miranda E., Granados-González G., de De la Torre L.G., Nishigaki T, & Darszon A. (2014). Zn2+ induces hyperpolarization by activation of a K+ channel and increases intracellular Ca2+ and pH in sea urchin spermatozoa. Developmental biology, 394(1), 15-23.
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3

Apoptosis Measurement by TUNEL

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1x104 621–101 cells/well were plated in 24 well plates, and serum depleted, or treated with NHLF conditioned serum free medium for 24 hours. Cells were fixed in 4% formaldehyde and the DNA fragmentation detected by TUNEL (Terminal deoxynucleotidyl transferase dUTP nick end labelling) using the Fluorescein In Situ Cell Death Detection Kit (Roche Diagnostics Limited, Burgess Hill, UK) according to manufacturer’s instructions. Cells were counterstained with 1ug/ml DAPI in PBS, imaged on a Nikon Diaphot 300 inverted microscope with epifluorescence under appropriate filters and the number of TUNEL (FITC) positive cells expressed as a percentage of the total number of DAPI stained nuclei.
Clements D., Dongre A., Krymskaya V.P, & Johnson S.R. (2015). Wild Type Mesenchymal Cells Contribute to the Lung Pathology of Lymphangioleiomyomatosis. PLoS ONE, 10(5), e0126025.
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